CD44v6 ELISA Kit

Produktnummer ABIN955787

Dieses Colorimetric ELISA kit dient der quantitativen Messung von Human CD44v6.

Kurzübersicht für CD44v6 ELISA Kit (ABIN955787)

Target: CD44v6

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Applikation: ELISA

Produktdetails

Analytische Methode: Quantitative

Spezifität: This assay recognizes both natural and recombinant forms of the sCD44var(v6) molecule. To define the specificity of this ELISA, several structurally related and non-related polypeptides were tested for cross reactivity. Notably, there was no detectable cross reactivity with CD44-polypeptides lacking the protein sequence encoded by exon 6.

Sensitivität: The limit of detection for sCD44var(v6), defined as the analyte concentration resulting in an absorption significantly higher than the absorption of the dilution medium (mean plus two standard deviations) was determined to be 0.05 ng/mL (mean of 10 independent assays).

Produktmerkmale: The Human sCD44var(v6) ELISA detects all circulating CD44 isoforms containing the sCD44var(v6) sequences. Determination of sCD44var(v6) will provide more detailed insight into different pathological modifications during cancer and other diseases.An anti-sCD44var(v6) monoclonal coating antibody is adsorbed onto microwells. sCD44var(v6) present in the sample or Calibrator binds to antibodies adsorbed to the microwells, a HRP-conjugated monoclonal anti-sCD44var(v6) antibody is added and binds to sCD44var(v6) captured by the first antibody. Following incubation unbound enzyme conjugated anti- sCD44var(v6) is removed during a wash step and substrate solution reactive with HRP is added to the wells. A colored product is formed in proportion to the amount of sCD44var(v6) present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450 nm. A calibration curve is prepared from six sCD44var(v6) Calibrator dilutions which allows determination of sCD44var(v6) concentration in samples.

Bestandteile: 1 aluminum pouch with a Microwell Plate coated with monoclonal antibody (mouse) to human sCD44var(v6)
2 vials (10 µL) HRP-Conjugate anti-sCD44var(v6) monoclonal (mouse) antibody
2 vials (Lyophilized) sCD44var(v6) Calibrator, 20 ng/mL upon reconstitution
1 bottle (50 mL) Wash Buffer Concentrate 20X, phosphate-buffered saline (PBS) with 1% Tween 20
1 vial (5 mL) Assay Buffer Concentrate 20X, PBS with 1% Tween 20 and protein stabilizer
1 bottle (50 mL) Sample Diluent (buffered protein matrix)
1 vial (7 mL) Substrate Solution I (tetramethylbenzidine)
1 vial (7 mL) Substrate Solution II (0.02% buffered hydrogen peroxide)
1 vial (12 mL) Stop Solution (1M Phosphoric acid)
1 vial (0.4 mL) Blue Dye
1 vial (0.4 mL) Green Dye
2 adhesive Plate Seals

Benötigtes Material: 5 mL and 10 mL graduated pipettes
5 µL to 1,000 µL adjustable single channel micropipettes with disposable tips
50 µL to 300 µL adjustable multichannel micropipette with disposable tips
Multichannel micropipette reservoir
Beakers, flasks, cylinders

Anwendungsinformationen

Plattentyp: Pre-coated

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: Since exact conditions may vary from assay to assay, a calibration curve must be established for every run.
Bacterial or fungal contamination of either screen samples or reagents or cross-contamination between reagents may cause erroneous results.

Lagerung: 4 °C

Informationen zur Lagerung: Store kit reagents at 4°C. Immediately after use reagents should be returned to cold storage (4°C). Expiration date of the kit and reagents is stated on labels. The expiration date of the kit components can only be guaranteed if the components are stored properly, and if, in case of repeated use of one component, the reagent is not contaminated by the first handling.

Haltbarkeit: The expiry date is stated on the label.

Antigendetails

Target: CD44v6

Andere Bezeichnung: SCD44var (v6)

Hintergrund: CD44 (Pgp-1, Ly-24, ECMR III, F10-44-2, H-CAM, HUTCH-I, In(Lu)-related p80, Hermes antigen, hyaluronan receptor) is a polymorphic glycoprotein which participates in a wide variety of cell-cell or cell-matrix interactions including lymphocyte homing, establishment of B and T cell immune responses, tumor metastasis formation and inflammation. Three isoform categories of the CD44 molecule have been identified: 1) a predominant 80-90 kDa category, the so-called standard form named C44std, 2) an intermediate size category of 110-160 kDa and 3) a category which includes very large isoforms of 250 kDa covalently modified by the addition of chondroitin sulfate. This CD44-family of transmembrane receptor molecules is derived from a single gene located on chromosome 11. Alternative splicing of the mRNA gives rise to the different isoforms, containing inserts of varying sizes in the extracellular domain of the molecule (exons v2-v10). All CD44 isoforms are variably glycosylated. In contrast to standard CD44 (CD44std) which is almost ubiquitously expressed, the variety of CD44 isoforms (CD44var) have a much more restricted distribution, e.g., on keratinocytes (exons v3-v10), epithelial cells (exons v8-v10), activated lymphocytes and macrophages (exon v6). A splice variant of CD44 (exons v4-v7) confers metastatic behavior in a rat carcinoma model, aberrant expression of splice variants has been detected on a variety of human tumor cell lines as well as primary and metastatic human tumors, including lymphomas, carcinomas (colon, thyroid, mamma, bladder), and gliomas.